ANIMAL MODEL OF HUMAN DISEASE
Bancroftian Filariasis
Wuchereria bancrofti Infection in the Silvered Leaf Monkey
(Presbytis cristatus)
JAMES R. PALMIERI, PhD (LT, MSC, USN),
DANIEL H. CONNOR, MD,
PURNOMO, DRS, and H. A. MARWOTO, DRA
Biologic Features
The silvered leaf monkey (Presbytis cristatus) is a
langur (Colobinae: Cercopithecidae) that inhabits
primary and secondary forest and mangrove swamp
ecotypes throughout Southeast Asia. P cristatus was
specifically chosen as a laboratory model of bancrof-
tian filariasis because of the recent finding in South
Kalimantan of Wuchereria kalimantani, a species
morphologically and taxonomically related to Wu-
chereria bancrofti. Because of this it was suspected
and subsequently shown that P cristatus was a suit-
able host for W bancrofti.
Most attempts to rear W bancrofti in a variety of
animals, including domestic cats,2'4 mongolian
jirds,5'6 hamsters,7 and monkeys,28-10 have failed to
obtain complete development of the worm. The two
most successful attempts were conducted by Cross et
al (1979)9 and Dissanaike and Mak (1980).10 In the
first study,9 exposure of 78 Macaca cyclopis monkeys
to third-stage larvae of W bancrofti (500-1550) re-
sulted in only 8 patent infections with low microfila-
remias (5-15/ml blood). In the second study,10 a
single immunosuppressed and splenectomized Ma-
caca fascicularis produced an occult infection con-
sisting of a single stunted adult W bancrofti. Recent-
0002-9440/83/0907-0383
383
From the Department of Infectious and Parasitic Disease Pathology,
Armed Forces Institute of Pathology, Washington, DC;
Department of Parasitology, US Naval Medical Research
Unit No. 2, Jakarta Detachment, APO San Francisco;
and Department of Parasitology, Indonesian Ministry
of Health Research and Development,
Jakarta, Indonesia
ly, Sucharit et al (1982)11 established W bancrofti in
a single immunosuppressed Presbytis melalophos.
In previous studies we inoculated infective third-
stage larvae of W bancrofti into 7 parasite-free
P cristatus.21-14 Each monkey received between 50
and 202 third-stage larvae of W bancrofti. Two
The opinions or assertions contained herein are those of
the authors and are not to be construed as official or as
reflecting the views of the Department of the Navy, Army,
or Defense or the Indonesian Ministry of Health.
The experiments reported herein were conducted accord-
ing to the principles set forth in the current edition of the
"Guide for the Care and Use of Laboratory Animals,"
prepared by the Committee on Care and Use of Laboratory
Animals of the Institute of Laboratory Animal Resources,
National Research Council.
Publication sponsored by the Registry of Comparative
Pathology of the Armed Forces Institute of Pathology and
supported by Public Health Service Grant RR-00301 from
the Division of Research Resources, National Institutes of
Health, under the auspices of Universities Associated for
Research and Education in Pathology, Inc.
Study supported by funds provided by the Indonesian
National Institutes of Health Research and Development;
the Naval Medical Research and Development Command,
Work Unit #ZF51.524.009.0085; and the WHO Collabo-
rating Center for Filarial Infections of Man, Armed Forces
Institute of Pathology.
 +*-O;NiQ&.xF%ag@:'Crj$|,Tw4sSKt_+P#va>
384 PALMIERI ET AL AJP * September 1983

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Vol. 112 * No. 3 BANCROFTIAN FILARIASIS 385

monkeys died 30 and 99 days later of unrelated
causes, 4 monkeys developed patent infections be-
tween 206 and 285 days, and 1 remained amicrofila-
remic after 400 days. Adult worms recovered from
the pericapsular region of the inguinal lymph node
and from the spermatic fascia of the testes and
microfilariae from venous blood were morphologi-
cally identical to parasites recovered from man
(Figure 1).
Comparison With Human Disease
The disease produced by W bancrofti in P cristatus
is indistinguishable from the human infection."1 At
necropsy, the two infected monkeys had lympha-
denopathy but no edema or other gross lesions.
Microscopically, the inguinal lymph nodes were con-
gested and had atrophic follicles, edema of the sub-
capsular sinusoids, and dilated lymphatics in both
cortex and medulla. The cortex contained many
plasma cells and eosinophils. In a dilated space in the
cortex there was an organized thrombus containing a
degenerating adult member of W bancrofti (Figures
6 and 7). Its cuticle was fragmented and surrounded
by foreign-body giant cells, and its internal organs
were replaced by inflammatory cells (Figures 8 and
9). Intact microfilariae were in the inguinal lymph
node of the monkey (Figure 2). A small focus of
lymphoid tissue in the perirenal fat also contained
microfilariae, and adjacent adipose tissue contained
a plexus of dilated lymphatics and blood vessels.
Eosinophils were in the intestinal mucosa and around
the bronchioles of the lung. There were discrete gran-
ulomas comprised of Langhans' giant cells sur-
rounded by eosinophils in the subcapsular region of
the spleen. Many of these giant cells contained de-
generating microfilariae (Figures 3-5). Otherwise the
spleen was diffusely congested and contained many
eosinophils and neutrophils.
In man, adult W bancrofti organisms invade and
distend lymphatic vessels and cause their walls to
thicken. Chronic bancroftian filariasis causes en-
largement of the lymph nodes, lymphedema, hydro-
cele, and sometimes elephantiasis. Dead and dying
worms form thrombi in the lymphatic vessels and are
surrounded by fibrin. Although the exact mechanism
of the lymphedema is not known, these inflammatory
changes retard and eventually obstruct the flow of
lymph. Chyluria and ascites may also develop. In
addition, bancroftian filariasis is probably the main
cause of tropical eosinophilia, including eosinophilic
lung and tropical pulmonary eosinophilia.1
Usefulness of the Model
The relationship of nonhuman primates to man is
important, especially in light of the evolutionary kin-
ship and the anatomic, physiologic, biochemical, and
behavioral similarities of the two species. It is not
surprising that the lesions caused by adult W ban-
crofti worms in P cristatus resemble those in man. P
cristatus was specifically inoculated with W bancrofti
larvae after one of us (J.R.P.) discovered and named
W kalimantani in wild P cristatus. This suggests a
long adaptive relationship between Wuchereria sp.
and the silvered leaf monkey. Until now, Wbancrofti
was thought to be specific for man, and experimental
infections had not produced a working model, the
lack of which has hindered our understanding of the
pathophysiology of filariasis as well as its treatment
and control. Filarial infections in P cristatus now
provide the opportunity for detailed study of the
interrelationships of host, parasite, and vector, all of
which may in turn lead to more effective treatment
and prevention. It is now feasible to study the pathol-
ogy and pathogenesis, host suitability, immunology,
and filaricidal drugs and vaccines.
Availability
P cristatus is common throughout the Malay
Archipelago but is especially plentiful in Malaysia
and Borneo. P cristatus is listed in Appendix II of the
Convention of International Trade of Endangered
Species and may be admitted into the United States.
Both the San Diego and the Los Angeles zoological
gardens have colonies of P cristatus. Work is now in
progress at the Armed Forces Institute of Pathology
to establish infections by Onchocerca volvulus and
Loa loa in P cristatus.

Figure 1-Microfilaria of Wuchereria bancrofti from the blood of an experimentally infected Presbytis cristatus (silvered leaf monkey). (Giemsa,
x 1000); AFIP 81-17386 Figure 2-Microfilariae (arrow) of Wuchereria bancrofti in inguinal lymph node. (H&E, x 630); AFIP 81-16890 Fig-
ure 3-Degenerating microfilariae of Wuchereria bancrofti in a giant cell from the spleen. Note the small filarial nuclei (arrow). (H&E, x 630);
AFIP 81-17383 Figure 4-Cross-section of a microfilaria (arrow) in a giant cell from the subcapsular region of the spleen. (Giemsa, x 1000);
AFIP 81-13960 Figure 5-Three sections of microfilariae (arrows) in a subcapsular giant cell of the spleen. (Epon/toluidine blue, x 1000);
AFIP 81-17384 Figure 6-Thrombus (arrow) in a lymphatic vessel of an inguinal lymph node. Note the edema in the subcapsular region
of the lymph node. (Movat, x 20); AFIP 81-17581 Figure 7-Higher magnification of Figure 6, showing giant cells surrounding degenerating
adult Wuchereria bancrofti. (Movat, x 60); AFIP 81-13945 Figure 8-Giant cell containing degenerating adult Wuchereria bancrofti and
infiltrated with host leukocytes. c, cuticle; g, giant cell. (H&E, x 250); AFIP 81-13946 Figure 9-Giant cell containing a cuticle from a degen-
erating adult Wuchereria bancrofti. c,cuticle; g, giant cell. (Movat, x250); AFIP 81-17577
386 PALMIERI ET AL
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AJP * September 1983
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Acknowledgments
We are indebted to Dr. Soekartono, Mr. Bertrand Ibra-
him, Mr. Suryatman Sudjar, Mr. Bambang Tenggono, and
Mr. Sofyan Masbar, who caught the monkeys and pro-
cessed the specimens; Mr. Aep Suhaepi, Mr. Nasir
Hamzah, and Mr. Soeroto Atmosoedjono, for studies of
the vectors, and Dr. Fauzi Darwis, for permission and sup-
port to carry out this study in the Banjar Regency, South
Kalimantan; and Capt. Elgin C. Cowart, Jr., Col. William
R. Cowan, and Maj. Adrian F. Van Dellen, of the Armed
Forces Institute of Pathology, Washington, DC, for con-
sultation reports and administrative support.